CN106479832B - Dabran koji fen-flavor liquor and production method thereof - Google Patents
Dabran koji fen-flavor liquor and production method thereof Download PDFInfo
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Abstract
The invention provides a production method of a bran koji fen-flavor liquor, which comprises the following steps: A) mixing the crushed sorghum, rice hulls, water and fermented grains, sequentially carrying out material sealing, material steaming, spreading and cooling, then inoculating bran koji, low-temperature Daqu, yeast and ester-producing yeast, and carrying out sealed fermentation to obtain fermented grains; B) and distilling the fermented grains and storing the distilled liquor to obtain the bran koji fen-flavor liquor. According to the method, the low-temperature Daqu, the bran koji, the ester-producing yeast and the yeast are used as saccharification leavening agents, and the sorghum is used as a fermentation raw material, so that the softness and the elegant degree of the base wine are improved, the total ester content in the base wine is increased, and the quality of the fen-flavor liquor is improved.
Description
Technical Field
The invention belongs to the technical field of white spirit, and particularly relates to a Dabran koji fen-flavor white spirit and a production method thereof.
Background
The white spirit yeast contains abundant microorganisms, and provides a mixed system of various microorganisms required by wine brewing. When the microorganism grows and breeds on the yeast block, various hydrolases are secreted, so that the yeast has liquefying power, saccharifying power, proteolytic power and the like. The Daqu contains multiple yeasts, and has fermenting and ester-producing effects. During the starter propagation process, the microorganisms decompose the raw materials to form metabolites, such as amino acids, ferulic acid, etc., which are precursors for forming the special flavor of the Daqu liquor, and the amino acids also provide the nitrogen source for brewing microorganisms.
The total ester is an important physicochemical index of the white spirit, and the esters in the white spirit are aromatic compounds, play an important role in various types of white spirits, are main factors for forming the main body style of the spirit body, and are important bases for dividing the flavor types of the white spirits.
The bran koji fen-flavor liquor is prepared by selecting sorghum as a raw material, taking bran koji and yeast as saccharification leavening agents, adopting a traditional 'five-old-rice-steamer' process, and performing the steps of raw material steaming, auxiliary material steaming, low-temperature tank entering, low-fire distillation, head and tail removing, storage and fine brewing. The liquor of the bran koji delicate fragrance type white spirit is clear and transparent, has fragrant fragrance, mellow wine quality, sweet and moist mouthfeel, is clear, has strong wine power, lasting aftertaste and long aftertaste.
With the development and progress of society, the dietary habits of consumers are developed from the past heavy taste to the light taste, the liquor industry is developing towards the direction of 'elegant and soft', the bran koji fen-flavor liquor has the taste advantages of fresh, pure and mellow taste, and the taste is not as soft as that of strong-flavor liquor and also not as fine as that of the Maotai-flavor liquor compared with the strong-flavor liquor and the Maotai-flavor liquor.
Disclosure of Invention
In view of the above, the technical problem to be solved by the invention is to provide the Dabran koji fen-flavor liquor and the preparation method thereof, and the Dabran koji fen-flavor liquor provided by the invention improves the softness and the elegant degree of the base liquor, increases the total ester content in the base liquor, and improves the quality of the fen-flavor liquor.
The invention provides a production method of a bran koji fen-flavor liquor, which comprises the following steps:
A) mixing the crushed sorghum, rice hulls, water and fermented grains, sequentially carrying out material sealing, material steaming, spreading and cooling, then inoculating bran koji, low-temperature Daqu, yeast and ester-producing yeast, and carrying out sealed fermentation to obtain fermented grains;
B) and distilling the fermented grains and storing the distilled liquor to obtain the bran koji fen-flavor liquor.
Preferably, the bran koji is prepared according to the following method:
inoculating Aspergillus niger HXM2 into wheat bran for fermentation to obtain bran koji;
the screening method of the Aspergillus niger HXM2 comprises the following steps:
adding the bacterial liquid containing the fen-flavor Daqu to a starch agar Chaochi culture medium for culturing, and screening to obtain Aspergillus niger HXM2 with the saccharifying enzyme activity of more than 3000 u/g.
Preferably, the low-temperature Daqu is prepared according to the following method:
making the barley and the peas into curved bricks;
naturally fermenting the bent brick to obtain low-temperature Daqu;
the natural fermentation process comprises the following steps: laying yeast, a mold feeding period, a mold airing period, a damp fire period, a strong fire period, a late fire period and a yeast culturing period, wherein the temperature of the strong fire period is 40-50 ℃.
Preferably, the yeast is selected from alcoholic yeast HXJ1, and the screening method of the alcoholic yeast HXJ1 comprises the following steps:
inoculating the fen-flavor Daqu in wort of 10-degree Brix, adding lactic acid, mixing, and culturing;
inoculating the culture into 10 ° Brix wort, adding lactic acid, mixing, and culturing;
repeating the steps for 3-4 times to obtain a yeast proliferation solution;
culturing the yeast proliferation solution on a wort agar culture medium of 10 DEG Brix, taking a typical colony, inoculating the typical colony on a test tube wort agar slant culture medium of 10 DEG Brix, and then sequentially inoculating the typical colony on a wort agar culture medium of 10 DEG Brix and a test tube wort agar slant culture medium of 10 DEG Brix for culturing to obtain the alcohol yeast HXJ1 with the alcohol production performance of more than 4.5% vol.
Preferably, the ester-producing yeast is selected from ester-producing yeast HXJ3, ester-producing yeast HXJ6 and ester-producing yeast HXJ7, and the screening method of the ester-producing yeast HXJ3, the ester-producing yeast HXJ6 and the ester-producing yeast HXJ7 comprises the following steps:
inoculating the fragrant yeast to wort of 10-degree Brix, adding lactic acid, mixing, and culturing;
inoculating the culture into 10 ° Brix wort, adding lactic acid, mixing, and culturing;
repeating the steps for 3-4 times to obtain a yeast proliferation solution;
after the yeast propagation liquid is placed on a wort agar culture medium with 10 degrees Brix and containing streptomycin for culture, a typical colony is taken and inoculated on a test tube wort agar slant culture medium with 10 degrees Brix, and then sequentially inoculated on the wort agar culture medium with 10 degrees Brix and the test tube wort agar slant culture medium with 10 degrees Brix for culture, and ester-producing yeast HXJ3 with the ester-producing property of 4.35g/L of ethyl acetate, ester-producing yeast HXJ6 with the ester-producing property of 4.20g/L of ethyl acetate and ester-producing yeast HXJ7 with the ester-producing property of 3.96g/L of ethyl acetate are respectively obtained.
Preferably, the ratio of the total inoculation amount of the bran koji, the low-temperature Daqu, the yeast and the ester-producing yeast to the using amount of the sorghum is (0.10-0.19): 1, the using amount ratio of the bran koji, the low-temperature Daqu, the yeast and the ester-producing yeast is (3-5): (5-8): (1-3): (1-3).
Preferably, the cellar entry temperature of the sealed fermentation is 14-18 ℃, the cellar entry water content of the sealed fermentation is 53-60%, the temperature of the sealed fermentation is 20-30 ℃, and the time of the sealed fermentation is 10-15 days.
Preferably, the distillation is carried out as follows:
and (3) uniformly and loosely paving a layer of 30-40 cm fermented grains on the slight strip of the steamer, introducing steam with the pressure of 0.02-0.05MPa, detecting the steam layer by layer, and feeding the steam into the steamer, wherein the liquor flowing speed of each steamer is less than 10 kg per minute, and the liquor flowing temperature is lower than 30 ℃, so that liquor is obtained.
Preferably, the storage time is not less than 120 days.
The invention also provides the bran koji fen-flavor liquor obtained by the production method.
Compared with the prior art, the invention provides a production method of the Dabran koji fen-flavor liquor, which comprises the following steps: A) mixing the crushed sorghum, rice hulls, water and fermented grains, sequentially carrying out material sealing, material steaming, spreading and cooling, then inoculating bran koji, low-temperature Daqu, yeast and ester-producing yeast, and carrying out sealed fermentation to obtain fermented grains; B) and distilling the fermented grains and storing the distilled liquor to obtain the bran koji fen-flavor liquor. According to the method, the low-temperature Daqu, the bran koji, the ester-producing yeast and the yeast are used as saccharification leavening agents, and the sorghum is used as a fermentation raw material, so that the softness and the elegant degree of the base wine are improved, the total ester content in the base wine is increased, and the quality of the fen-flavor liquor is improved.
Detailed Description
The invention provides a production method of a bran koji fen-flavor liquor, which comprises the following steps:
A) mixing the crushed sorghum, rice hulls, water and fermented grains, sequentially carrying out material sealing, material steaming, spreading and cooling, then inoculating bran koji, low-temperature Daqu, yeast and ester-producing yeast, and carrying out sealed fermentation to obtain fermented grains;
B) and distilling the fermented grains and storing the distilled liquor to obtain the bran koji fen-flavor liquor.
Firstly, mixing the crushed sorghum, the rice hulls, water and the fermented grains to obtain a mixture.
Wherein the fermented grains are fermented grains obtained by distilling wine. The mass ratio of the sorghum, the rice hull, the water and the fermented grains is preferably 1 (0.2-0.3) to 0.5-0.8 to 3.8-4.5.
Then, the invention carries out material sealing, material steaming, spreading and cooling on the mixture.
The temperature of the material sealing is preferably 60-70 ℃, and the time of the material sealing is more than or equal to 30 min.
And after the material stewing is finished, steaming the material, wherein the specific method for steaming the material comprises the following steps:
and (3) lightly sprinkling the mixture, filling the mixture into a retort barrel, boiling the mixture for gelatinization after steaming, and counting more than 40 minutes from the round steam. The mixture is required to be completely gelatinized, cooked but not sticky, and has no core.
And spreading and cooling the mixture after steaming until the mixture is cooled to 30-35 ℃.
Then inoculating bran koji, low-temperature Daqu, yeast and ester-producing yeast to the mixture, and performing sealed fermentation to obtain fermented grains;
wherein the bran koji is prepared by the following method:
inoculating Aspergillus niger HXM2 into wheat bran for fermentation to obtain bran koji;
the Aspergillus niger HXM2 is purchased from Beijing Red Star GmbH, and the screening method comprises the following steps:
adding the bacterial liquid containing the fragrant yeast into a starch agar Chaochou culture medium for culturing, and screening to obtain Aspergillus niger HXM2 with the saccharifying enzyme activity of more than 3000 u/g.
Specifically, the invention takes the fragrant yeast as a separation source, and the invention has no special limitation on the preparation method of the fragrant yeast, and the preparation method of the fragrant yeast is known by the technical personnel in the field.
Taking a starch agar Czochralski culture medium as an isolation culture medium, wherein the starch agar Czochralski culture medium comprises: starch 2%, NaNO30.3％，KCl 0.05％，K2HPO40.1％，FeSO40.001％，MgSO40.5%, agar 2%, and water in balance, pH6.7, and sterilizing at 121 deg.C for 20 min.
Separating agent: 0.02mol/L iodine solution and sterile water.
And (3) a separation process: heating and melting starch agar Czochralski culture medium, pouring 12-15ml of the starch agar Czochralski culture medium into each of 3 sterile culture dishes, and uniformly distributing by rotation. Adding a small amount of fragrant Daqu powder into a small triangular flask with a volume of 20ml containing sterile water and glass beads, shaking vigorously to break up spore granules, and filtering with several layers of sterile gauze in 1 sterile test tube. Diluting the filtrate with sterile water to 10 deg.C-1、10-2、10-3Three fold dilutions. 0.15ml of the bacteria-containing solution with 3 dilutions is respectively added on a starch agar Chao's medium plate, after being evenly coated by a coating rod, the culture dish is placed in a thermostat with the temperature of 30 ℃ for culture. When the bacterial colony is preliminarily formed and no spore is produced, iodine solution is dripped around the bacterial colony, the bacterial colony with a large transparent ring is selected and transferred to the inclined plane of a starch agar Cnahs medium, the temperature is kept at 30 ℃, after about 5 days of culture, the shape of each purified bacterial strain is observed, and the activity performance of the glucoamylase of each bacterial strain is measured.
And (3) measuring the activity performance of the saccharifying enzyme: the strain is selected and inoculated into a bran solid culture medium (bran: water 1:0.9) sterilized at 121 ℃ for 30 minutes, cultured in a constant temperature box at 30 ℃ for 5 days, and the culture is subjected to diastase activity determination according to QB1805.2-1993 industrial diastase preparations.
After multiple screening, Aspergillus niger HXM2 with saccharifying enzyme activity of more than 3000u/g is screened.
In the present invention, the low-temperature yeast is preferably prepared as follows:
making the barley and the peas into curved bricks;
naturally fermenting the bent brick to obtain low-temperature Daqu;
the natural fermentation process comprises the following steps: laying yeast, a mold feeding period, a mold airing period, a damp fire period, a strong fire period, a late fire period and a yeast culturing period, wherein the temperature of the strong fire period is 40-50 ℃.
Specifically, the fen-flavor Daqu is prepared by firstly making barley and peas (6:4) into a yeast brick and naturally fermenting in a yeast room.
The fermentation temperature is controlled by opening and closing a window by a master of a koji house during the fermentation process.
The fermentation culture process comprises a yeast laying period, a mold feeding period, a mold airing period, a damp fire period, a strong fire period, a late fire period and a yeast raising period.
① laying the yeast, laying the yeast blank of the fen-flavor Daqu in a room, spreading the yeast blank with dry bran, laying the yeast blank in an upper layer and a lower layer, and making the yeast blank with the distance of 3-4 cm between the reed stalks, wherein the yeast blank is arranged in a row after another row without the distance of rows.
②, getting mildewed, adjusting the temperature of the yeast chamber to a certain temperature after the yeast blank is put into a room, 12-15 ℃ in winter, 15-18 ℃ in spring and autumn, and keeping the temperature as far as possible in summer, after the surfaces of the yeast blocks are dried in the air, spraying little cold water with a spray can to cover the reed mat, spraying water to moisten the reed mat, slowly heating the reed mat, slowly igniting, controlling the temperature in winter to be 72-80 hours, raising the temperature of the yeast to 38 ℃, getting mildewed well, and in summer, raising the temperature rapidly due to high temperature, and reaching 38 ℃ only after 38-40 hours.
③ air drying for mould, opening the reed mat when the mould on the surface of the yeast blank is good, opening the window to release damp, drying the yeast skin, turning over the yeast for the first time, turning over the three layers into four layers, separating the middle by the reed mat, arranging the four layers in a shape like a Chinese character 'pin', turning over the yeast at the yeast interval of 3-4 cm., closing the window to fire at 28-32 ℃, setting two times at day and night, sealing the two times at the window, heating the yeast to 32-36 ℃, turning over the yeast blocks from four layers into five layers, and the arrangement method is the same.
④, starting damp fire, preparing in advance for 1 day, heating the product in the middle of the koji mold to 38 ℃ just, when the koji mold is turned into six layers from five layers, extracting reed rods, arranging the koji mold into a herringbone shape, turning into seven layers from six layers after 1 day, leaving a flame path in the middle, heating up two times and two drops in the day and night, sealing two times in the window, gradually heating up 4-5 days, cooling down the koji mold when the top temperature reaches 44-46 ℃, cooling down the koji mold (experience), and taking the temperature of the koji mold to be reduced to 28-30 ℃ as a limit.
⑤ strong fire, the temperature from the damp fire to the top of the hot yeast is 40-50 ℃, more preferably 44-46 ℃, the strong fire period begins, the temperature rises twice in day and night, the temperature rises twice in window, the temperature is kept at 44-46 ℃ in about 7-8 days, the temperature of the top of the hot yeast is kept at 28-30 ℃, and the yeast is turned over once a day in the strong fire period.
⑥ after fire, gradually decreasing from the top temperature of the big fire hot yeast to 44-46 ℃ until the yeast block is hot, but the yeast core still has residual heat, then adding external heat, keeping the top temperature of the hot yeast to be 32-33 ℃, cooling the yeast to be 28-30 ℃ in the air, needing 5-6 days approximately, and turning over the yeast once every 2-3 days.
⑦ starter culture, slightly residual heat in the starter core, heating to keep the starter block at 32-33 ℃, cooling the starter to 28-30 ℃, maintaining for 3-4 days, and culturing for 24-25 days in total, which is no more than 28 days.
⑧ when the starter culture period is over, only a little water in the starter core is removed, and the starter block can be taken out.
In the invention, the yeast is selected from alcoholic yeast HXJ1, purchased from Beijing Hongxing GmbH, and the screening method of the alcoholic yeast HXJ1 is as follows:
inoculating the fragrant yeast to wort of 10-degree Brix, adding lactic acid, mixing, and culturing;
inoculating the culture into 10 ° Brix wort, adding lactic acid, mixing, and culturing;
repeating the steps for 3-4 times to obtain a yeast proliferation solution;
culturing the yeast proliferation solution on a wort agar culture medium of 10 DEG Brix, taking a typical colony, inoculating the typical colony again and culturing on the wort agar culture medium of 10 DEG Brix to obtain alcoholic yeast HXJ1 with the alcohol production performance of more than 4.5% vol.
The method takes the fragrant yeast as a separation source, and sequentially performs enrichment culture and plate separation to obtain the alcohol yeast HXJ 1.
Specifically, the enrichment culture method comprises the following steps:
a sample of rice size was scooped up from the desired portion of the koji block with a sterile knife, added to a sterile 10ml wort test tube of 10 ° Brix, and 1 drop of lactic acid was added thereto, shaken well, and then cultured in a thermostat at 25 ℃ for 24 hours. 1ml of the above culture was transferred to another 1 sterile malt wort lactic acid test tube of 10 ℃ Brix and cultured again. After 3-4 times of transferring and culturing, plate separation can be carried out.
Plate separation: taking 1ml of the final 1 generation yeast proliferation solution, diluting with sterile water to 10%-1、10-2、10-3Three fold dilutions. 0.15ml of yeast liquid with 3 dilutions was taken and added to a sterile 10 ° Brix malt agar medium plate, and after coating evenly with a coating rod, the plate was placed upside down in a thermostat at 25 ℃ for 48 hours. Typical colonies are picked from the plate and transferred to malt extract agar slant culture medium of 10 ° Brix test tube, and after culturing at 25 ℃, the plate is separated again and transferred to the test tube slant for morphological and physiological performance research.
And (3) alcohol production performance determination: the fermentation medium is sorghum saccharified solution of 12 ° Brix, and is sterilized at 115 deg.C for 20 min. The test tube strain is selected from a ring fungus and inoculated into a 20ml sorghum saccharification liquid test tube, and cultured for 24 hours at 28 ℃. The cultured test tube seeds are inoculated into 200ml of sorghum saccharification liquid and fermented for 3 days at 28 ℃. Distilling the fermented fermentation liquor, and measuring the alcoholic strength of the distillate by using an alcohol meter.
After a plurality of screening, the alcohol yeast HXJ1 with the alcohol production performance of more than 4.5 percent vol is screened.
In the invention, the liquor culture process of the yeast wine comprises the following steps: adding 10 g of glucose and 150 g of distilled grain 100-one into a certain amount of water, uniformly mixing, sterilizing for 20 minutes at 0.1MPa, and culturing for 12-16 hours at 30-32 ℃ after inoculation.
The vinasse is a substance obtained by distilling fermented grains.
In the invention, the ester-producing yeast is selected from ester-producing yeast HXJ3, ester-producing yeast HXJ6 and ester-producing yeast HXJ7, which are all purchased from Beijing Hongxing GmbH, and the screening method of the ester-producing yeast HXJ3, the ester-producing yeast HXJ6 and the ester-producing yeast HXJ7 comprises the following steps:
inoculating the fragrant yeast to wort of 10-degree Brix, adding lactic acid, mixing, and culturing;
inoculating the culture into 10 ° Brix wort, adding lactic acid, mixing, and culturing;
repeating the steps for 3-4 times to obtain a yeast proliferation solution;
after the yeast proliferation solution is placed on a wort agar culture medium containing streptomycin and having a Brix of 10 degrees for culture, a typical colony is taken and inoculated on the wort agar culture medium having a Brix of 10 degrees for culture again, and ester-producing yeast HXJ3 with the ester-producing property of 4.35g/L of ethyl acetate content, ester-producing yeast HXJ6 with the ester-producing property of 4.20g/L of ethyl acetate content and ester-producing yeast HXJ7 with the ester-producing property of 3.96g/L of ethyl acetate content are respectively obtained.
The invention takes the fragrant yeast as a separation source to carry out enrichment culture and plate separation in turn.
Wherein, the culture medium: wort of 10 ° Brix is used as enrichment medium, and wort agar of 10 ° Brix is used as isolation medium.
Separating agent: lactic acid, streptomycin, sterile water.
The specific method of enrichment culture comprises the following steps:
a sample of rice size is dug out from the required part of the fragrant Daqu yeast block by a sterile knife, added into a sterile 10ml malt wort test tube, added with 1 drop of lactic acid, shaken up and cultured in a thermostat at 25 ℃ for 24 hours. 1ml of the above culture was transferred to another 1 sterile malt wort lactic acid test tube and cultured again. After 3-4 times of transferring and culturing, plate separation can be carried out.
The specific method for separating the flat plate comprises the following steps:
taking 1ml of the final 1 generation yeast ester-increasing solution, diluting with sterile water to 10%-1、10-2、10-3Three fold dilutions. 0.15ml of yeast liquid of 3 dilutions was added to sterile malt extract agar medium plates of 10 ° Brix of 30ug/ml streptomycin, and after coating evenly with a coating rod, the plates were placed upside down in a thermostat at 25 ℃ for 48 hours. Typical colonies picked on the plates were transferred to test tube wort agar slants of 10 ° BrixAfter culturing at 25 ℃ in a surface culture medium, the medium was again inoculated into a wort agar medium of 10 ° Brix for plate separation, and transferred to a test tube wort agar slant medium of 10 ° Brix for morphological and physiological performance studies.
And (3) measuring the ester production performance: the fermentation medium is sorghum saccharified solution of 12 ° Brix, and is sterilized at 115 deg.C for 20 min. The test tube strain is selected from a ring fungus and inoculated into a 20ml sorghum saccharification liquid test tube, and cultured for 24 hours at 28 ℃. The cultured test tube seeds are inoculated into 200ml of sorghum saccharification liquid and fermented for 4 days at 28 ℃. Distilling the fermented fermentation liquor, and analyzing the ethyl acetate content of the distillate by gas chromatography.
Through multiple screening, the ester-producing yeast HXJ3 with the ester-producing performance of 4.35g/L of ethyl acetate, the ester-producing yeast HXJ6 with the ester-producing performance of 4.20g/L of ethyl acetate and the ester-producing yeast HXJ7 with the ester-producing performance of 3.96g/L of ethyl acetate are obtained respectively.
In the invention, the liquid culture process of the ester-producing yeast comprises the following steps: adding 10 g of glucose and 100-150 g of vinasse into a certain amount of water, uniformly mixing, sterilizing at 0.1MPa for 20 minutes, and culturing at 25-28 ℃ for 16-20 hours after inoculation.
In the invention, the ratio of the total inoculation amount of the bran koji, the low-temperature Daqu, the yeast and the ester-producing yeast to the using amount of the sorghum is (0.10-0.19): 1, preferably (0.12-0.17): 1.
the using amount ratio of the bran koji, the low-temperature Daqu, the yeast and the ester-producing yeast is (3-5): (5-8): (1-3): (1-3), preferably (3.5-4.5): (6-7): (1.5-2.5): (1.5-2.5).
The inoculated mixture is subjected to sealed fermentation, and the cellaring temperature of the sealed fermentation is 14-18 ℃, preferably 15-17 ℃; the cellar entry water content of the sealed fermentation is 53-60%, and preferably 55-58%; the temperature of the sealed fermentation is 20-30 ℃, and preferably 23-28 ℃; the time for sealed fermentation is 10-15 days, and preferably 12-14 days.
And after the fermentation is finished, obtaining fermented grains. The fermented grains are distilled and stored to obtain the bran koji fen-flavor liquor.
In the present invention, the distillation is preferably carried out as follows:
and (3) uniformly and loosely paving a layer of 30-40 cm fermented grains on the slight strip of the steamer, introducing steam with the pressure of 0.02-0.05MPa, detecting the steam layer by layer, and feeding the steam into the steamer, wherein the liquor flowing speed of each steamer is less than 10 kg per minute, and the liquor flowing temperature is lower than 30 ℃, so that liquor is obtained.
And finally, storing the obtained wine liquid, wherein the storage time is not less than 120 days, and the storage temperature is room temperature.
The invention also provides the bran koji fen-flavor liquor obtained by the production method.
According to the method, the low-temperature Daqu, the bran koji, the ester-producing yeast and the yeast are used as saccharification leavening agents, and the sorghum is used as a fermentation raw material, so that the softness and the elegant degree of the base wine are improved, the total ester content in the base wine is increased, and the quality of the fen-flavor liquor is improved.
In order to further understand the present invention, the following examples are provided to illustrate the bran koji fen-flavor liquor and the production method thereof, and the scope of the present invention is not limited by the following examples.
Example 1 preparation of bran koji
(1) Screening of Aspergillus niger HXM2
The method takes the fragrant yeast as a separation source and takes a starch agar Chao's medium as a separation medium, wherein the starch agar Chao's medium comprises the following components: starch 2%, NaNO30.3％，KCl 0.05％，K2HPO40.1％，FeSO40.001％，MgSO40.5%, agar 2%, and water in balance, pH6.7, and sterilizing at 121 deg.C for 20 min.
Separating agent: 0.02mol/L iodine solution and sterile water.
And (3) a separation process: heating and melting starch agar Czochralski culture medium, pouring 12-15ml of the starch agar Czochralski culture medium into each of 3 sterile culture dishes, and uniformly distributing by rotation. Adding a small amount of fragrant Daqu powder into a small triangular flask with a volume of 20ml containing sterile water and glass beads, shaking vigorously to break up spore granules, and filtering with several layers of sterile gauze in 1 sterile test tube. Diluting the filtrate with sterile water to 10 deg.C-1、10-2、10-3Three fold dilutions. 0.15ml of the bacteria-containing solution with 3 dilutions is respectively added on a starch agar Chao's medium plate, after being evenly coated by a coating rod, the culture dish is placed in a thermostat with the temperature of 30 ℃ for culture. When the bacterial colony is preliminarily formed and no spore is produced, iodine solution is dripped around the bacterial colony, the bacterial colony with a large transparent ring is selected and transferred to the inclined plane of a starch agar Cnahs medium, the temperature is kept at 30 ℃, after about 5 days of culture, the shape of each purified bacterial strain is observed, and the activity performance of the glucoamylase of each bacterial strain is measured.
And (3) measuring the activity performance of the saccharifying enzyme: the strain is selected and inoculated into a bran solid culture medium (bran: water 1:0.9) sterilized at 121 ℃ for 30 minutes, cultured in a constant temperature box at 30 ℃ for 5 days, and the culture is subjected to diastase activity determination according to QB1805.2-1993 industrial diastase preparations.
After multiple screening, Aspergillus niger HXM2 with saccharifying enzyme activity of more than 3000u/g is screened.
(2) Preparation of bran koji
Aspergillus niger HXM2 is inoculated into wheat bran for fermentation to obtain bran koji.
Example 2 preparation of Low temperature Daqu
Firstly, making the barley and the peas (6:4) into a yeast brick, and naturally fermenting in a yeast room.
The fermentation temperature is controlled by opening and closing a window by a master of a koji house during the fermentation process.
The fermentation culture process comprises a yeast laying period, a mold feeding period, a mold airing period, a damp fire period, a strong fire period, a late fire period and a yeast raising period.
① laying the yeast, laying the yeast blank of the fen-flavor Daqu in a room, spreading the yeast blank with dry bran, laying the yeast blank in an upper layer and a lower layer, and making the yeast blank with the distance of 3-4 cm between the reed stalks, wherein the yeast blank is arranged in a row after another row without the distance of rows.
②, getting mildewed, adjusting the temperature of the yeast chamber to a certain temperature after the yeast blank is put into a room, 12-15 ℃ in winter, 15-18 ℃ in spring and autumn, and keeping the temperature as far as possible in summer, after the surfaces of the yeast blocks are dried in the air, spraying little cold water with a spray can to cover the reed mat, spraying water to moisten the reed mat, slowly heating the reed mat, slowly igniting, controlling the temperature in winter to be 72-80 hours, raising the temperature of the yeast to 38 ℃, getting mildewed well, and in summer, raising the temperature rapidly due to high temperature, and reaching 38 ℃ only after 38-40 hours.
③ air drying for mould, opening the reed mat when the mould on the surface of the yeast blank is good, opening the window to release damp, drying the yeast skin, turning over the yeast for the first time, turning over the three layers into four layers, separating the middle by the reed mat, arranging the four layers in a shape like a Chinese character 'pin', turning over the yeast at the yeast interval of 3-4 cm., closing the window to fire at 28-32 ℃, setting two times at day and night, sealing the two times at the window, heating the yeast to 32-36 ℃, turning over the yeast blocks from four layers into five layers, and the arrangement method is the same.
④, starting damp fire, preparing in advance for 1 day, heating the product in the middle of the koji mold to 38 ℃ just, when the koji mold is turned into six layers from five layers, extracting reed rods, arranging the koji mold into a herringbone shape, turning into seven layers from six layers after 1 day, leaving a flame path in the middle, heating up two times and two drops in the day and night, sealing two times in the window, gradually heating up 4-5 days, cooling down the koji mold when the top temperature reaches 44-46 ℃, cooling down the koji mold (experience), and taking the temperature of the koji mold to be reduced to 28-30 ℃ as a limit.
⑤ strong fire, the temperature from the damp fire to the top of the hot yeast is 44-46 ℃, the strong fire period begins, the temperature rises twice in day and night, the temperature rises twice in windows, the temperature is kept at 44-46 ℃ in about 7-8 days, the temperature of the top of the hot yeast is kept at 28-30 ℃, and the yeast is turned over once every day in the strong fire period.
⑥ after fire, gradually decreasing from the top temperature of the big fire hot yeast to 44-46 ℃ until the yeast block is hot, but the yeast core still has residual heat, then adding external heat, keeping the top temperature of the hot yeast to be 32-33 ℃, cooling the yeast to be 28-30 ℃ in the air, needing 5-6 days approximately, and turning over the yeast once every 2-3 days.
⑦ starter culture, slightly residual heat in the starter core, heating to keep the starter block at 32-33 ℃, cooling the starter to 28-30 ℃, maintaining for 3-4 days, and culturing for 24-25 days in total, which is no more than 28 days.
⑧ when the starter culture period is over, only a little water in the starter core is removed, and the starter block can be taken out.
Example 3
A sample of rice size was scooped up from the desired portion of the koji block with a sterile knife, added to a sterile 10ml wort test tube of 10 ° Brix, and 1 drop of lactic acid was added thereto, shaken well, and then cultured in a thermostat at 25 ℃ for 24 hours. 1ml of the above culture was transferred to another 1 sterile malt wort lactic acid test tube of 10 ℃ Brix and cultured again. After 3-4 times of transferring and culturing, plate separation can be carried out.
Plate separation: taking 1ml of the final 1 generation yeast proliferation solution, diluting with sterile water to 10%-1、10-2、10-3Three fold dilutions. 0.15ml of yeast liquid with 3 dilutions was taken and added to a sterile 10 ° Brix malt agar medium plate, and after coating evenly with a coating rod, the plate was placed upside down in a thermostat at 25 ℃ for 48 hours. Typical colonies are picked from the plate and transferred to malt extract agar slant culture medium of 10 ° Brix test tube, and after culturing at 25 ℃, the plate is separated again and transferred to the test tube slant for morphological and physiological performance research.
And (3) alcohol production performance determination: the fermentation medium is sorghum saccharified solution of 12 ° Brix, and is sterilized at 115 deg.C for 20 min. The test tube strain is selected from a ring fungus and inoculated into a 20ml sorghum saccharification liquid test tube, and cultured for 24 hours at 28 ℃. The cultured test tube seeds are inoculated into 200ml of sorghum saccharification liquid and fermented for 3 days at 28 ℃. Distilling the fermented fermentation liquor, and measuring the alcoholic strength of the distillate by using an alcohol meter.
After a plurality of screening, the alcohol yeast HXJ1 with the alcohol production performance of more than 4.5 percent vol is screened.
Example 4
A sample of rice size is dug out from the required part of the fragrant Daqu yeast block by a sterile knife, added into a sterile 10ml malt wort test tube, added with 1 drop of lactic acid, shaken up and cultured in a thermostat at 25 ℃ for 24 hours. 1ml of the above culture was transferred to another 1 sterile malt wort lactic acid test tube and cultured again. After 3-4 times of transferring and culturing, plate separation can be carried out.
The specific method for separating the flat plate comprises the following steps:
taking the last generation yeast 1 to increase the ester content1ml, diluted to 10 with sterile water-1、10-2、10-3Three fold dilutions. 0.15ml of yeast liquid of 3 dilutions was added to sterile malt extract agar medium plates of 10 ° Brix of 30ug/ml streptomycin, and after coating evenly with a coating rod, the plates were placed upside down in a thermostat at 25 ℃ for 48 hours. Typical colonies were picked on plates and transferred to a test tube wort agar slant medium of 10 ° Brix for culture at 25 ℃, and then again inoculated to a 10 ° Brix wort agar slant medium for plate separation, and transferred to a test tube wort agar slant medium of 10 ° Brix for morphological and physiological performance studies.
And (3) measuring the ester production performance: the fermentation medium is sorghum saccharified solution of 12 ° Brix, and is sterilized at 115 deg.C for 20 min. The test tube strain is selected from a ring fungus and inoculated into a 20ml sorghum saccharification liquid test tube, and cultured for 24 hours at 28 ℃. The cultured test tube seeds are inoculated into 200ml of sorghum saccharification liquid and fermented for 4 days at 28 ℃. Distilling the fermented fermentation liquor, and analyzing the ethyl acetate content of the distillate by gas chromatography.
Through multiple screening, the ester-producing yeast HXJ3 with the ester-producing performance of 4.35g/L of ethyl acetate, the ester-producing yeast HXJ6 with the ester-producing performance of 4.20g/L of ethyl acetate and the ester-producing yeast HXJ7 with the ester-producing performance of 3.96g/L of ethyl acetate are obtained respectively.
Example 5
1000 kg of crushed sorghum, adding rice husks, water and 4000 kg of fermented grains, steaming for 35 min, steaming for 40 min, spreading and cooling to 30-35 ℃, inoculating 35 kg of bran koji obtained in example 1, 80 kg of low-temperature Daqu obtained in example 2, 25 kg of yeast obtained in example 3, 30 kg of ester-producing yeast obtained in example 4, placing the materials into a cellar at 15 ℃, placing the materials into the cellar for 55.3% of water, turning and stirring the materials into the cellar for treading tightly, sealing and fermenting for 15 days, and then taking the materials out of the cellar to obtain fermented grains.
And (3) uniformly and loosely laying a layer of 30-40 cm fermented grains on the slight strip of the retort, introducing steam with the pressure of 0.02-0.05MPa, detecting the steam layer by layer, loading the steam into the retort, wherein the liquor flowing speed of each retort is less than 10 kg per minute, the liquor flowing temperature is lower than 30 ℃, and the liquor obtained after distillation is stored for four months for use.
Example 6
Adding 1000 kg of crushed sorghum into rice husks, water and 4500 kg of fermented grains, sealing the materials for 30min, steaming the materials for 45 min, spreading and cooling the materials to 30-35 ℃, inoculating 50 kg of bran koji obtained in example 1, 55 kg of low-temperature Daqu obtained in example 2, 30 kg of yeast obtained in example 3, 20 kg of ester-producing yeast obtained in example 4, placing the materials into a cellar for 18 ℃, placing the materials into the cellar for 57.6% of water, turning the materials into the cellar, treading the materials tightly, sealing and fermenting the materials for 10 days, and then taking the materials out of the cellar to obtain fermented grains.
And (3) uniformly and loosely laying a layer of 30-40 cm fermented grains on the slight strip of the retort, introducing steam with the pressure of 0.02-0.05MPa, detecting the steam layer by layer, loading the steam into the retort, wherein the liquor flowing speed of each retort is less than 10 kg per minute, the liquor flowing temperature is lower than 30 ℃, and the liquor obtained after distillation is stored for four months for use.
Example 7
Adding 1000 kg of crushed sorghum into rice husks, water and distilled liquor, then adding 4000 kg of fermented grains, stewing for 40 min, steaming for 40 min, spreading and cooling to 30-35 ℃, inoculating 40 kg of bran koji obtained in example 1, 75 kg of low-temperature Daqu obtained in example 2, 30 kg of yeast obtained in example 3 and 25 kg of ester-producing yeast obtained in example 4, placing the materials into a cellar for 16.5 ℃, placing the materials into the cellar for 56.9% of water, turning and stirring the materials into the cellar for tight fermentation for 12 days, and then taking the materials out of the cellar to obtain fermented grains.
And (3) uniformly and loosely laying a layer of 30-40 cm fermented grains on the slight strip of the retort, introducing steam with the pressure of 0.02-0.05MPa, detecting the steam layer by layer, loading the steam into the retort, wherein the liquor flowing speed of each retort is less than 10 kg per minute, the liquor flowing temperature is lower than 30 ℃, and the liquor obtained after distillation is stored for four months for use.
Control 1:
adding 1000 kg of crushed sorghum into rice husks, water and distilled liquor, then returning 4000 kg of fermented grains to be subjected to material sealing for 35 min, steaming the materials for 40 min, spreading and cooling to 30-35 ℃, inoculating 80 kg of bran koji prepared in example 1 and 40 kg of yeast prepared in example 3, placing the materials into a cellar at the temperature of 18 ℃, placing the materials into the cellar with the moisture of 58.1%, turning and placing the materials into the cellar to be tightly treaded, sealing and fermenting for 8 days, and then taking the materials out of the cellar to obtain fermented grains.
And (3) uniformly and loosely laying a layer of 30-40 cm fermented grains on the slight strip of the retort, introducing steam with the pressure of 0.02-0.05MPa, detecting the steam layer by layer, loading the steam into the retort, wherein the liquor flowing speed of each retort is less than 10 kg per minute, the liquor flowing temperature is lower than 30 ℃, and the liquor obtained after distillation is stored for four months for use.
Control 2:
adding 1000 kg of crushed sorghum into rice husks, water and distilled liquor, then returning 4200 kg of fermented grains to be subjected to material sealing for 35 min, steaming the materials for 40 min, spreading and cooling to 30-35 ℃, inoculating 80 kg of bran koji prepared in example 1 and 40 kg of yeast prepared in example 3, placing the materials into a cellar at the temperature of 17 ℃, placing the materials into the cellar for 56.8% of water, turning and placing the materials into the cellar for treading tightly, sealing and fermenting for 8 days, and then taking the materials out of the cellar to obtain fermented grains.
And (3) uniformly and loosely laying a layer of 30-40 cm fermented grains on the slight strip of the retort, introducing steam with the pressure of 0.02-0.05MPa, detecting the steam layer by layer, loading the steam into the retort, wherein the liquor flowing speed of each retort is less than 10 kg per minute, the liquor flowing temperature is lower than 30 ℃, and the liquor obtained after distillation is stored for four months for use.
Control 3:
adding 1000 kg of crushed sorghum into rice husks, water and distilled liquor, then returning 4000 kg of fermented grains to be subjected to material sealing for 35 min, steaming the materials for 40 min, spreading and cooling to 30-35 ℃, inoculating 80 kg of bran koji prepared in example 1 and 40 kg of yeast prepared in example 3, placing the materials into a cellar at the temperature of 16.5 ℃, placing the materials into the cellar with the moisture of 57.5%, turning and stirring the materials into the cellar to be tightly treaded, sealing and fermenting for 8 days, and then taking the materials out of the cellar to obtain fermented grains.
And (3) uniformly and loosely laying a layer of 30-40 cm fermented grains on the slight strip of the retort, introducing steam with the pressure of 0.02-0.05MPa, detecting the steam layer by layer, loading the steam into the retort, wherein the liquor flowing speed of each retort is less than 10 kg per minute, the liquor flowing temperature is lower than 30 ℃, and the liquor obtained after distillation is stored for four months for use.
Example 8
And (3) measuring the total ester content in the white spirit by adopting an acid-base neutralization titration method. The content of ethyl acetate and ethyl lactate in the white spirit is measured by gas chromatography. The results are shown in Table 1.
TABLE 1 determination of the ester content of white spirit
| Total esters (g/l) | Ethyl acetate (g/l) | Ethyl lactate (g/l) | |
| Example 1 | 2.93 | 1.75 | 1.26 |
| Example 2 | 2.09 | 1.19 | 1.03 |
| Example 3 | 2.64 | 1.51 | 1.16 |
| Control 1 | 1.49 | 0.81 | 0.71 |
| Control 2 | 1.23 | 0.70 | 0.61 |
| Control 3 | 1.40 | 0.83 | 0.48 |
As can be seen from Table 1, the total esters of the examples are greatly improved over the total esters of the control.
Example 9
The results of sensory evaluation of the white spirits obtained in the above examples and controls by 30 professional tasters were averaged and are shown in table 2.
TABLE 2 sensory evaluation results of white spirit
As can be seen from Table 2, the white spirit provided by the embodiment improves the softness and the elegant degree of the base spirit, increases the total ester content in the base spirit, and improves the quality of the fen-flavor white spirit.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (6)
1. The production method of the bran koji fen-flavor liquor is characterized by comprising the following steps:
A) mixing the crushed sorghum, rice hulls, water and fermented grains, sequentially carrying out material sealing, material steaming, spreading and cooling, then inoculating bran koji, low-temperature Daqu, yeast and ester-producing yeast, and carrying out sealed fermentation to obtain fermented grains;
the bran koji is prepared according to the following method:
inoculating Aspergillus niger HXM2 into wheat bran for fermentation to obtain bran koji;
the screening method of the Aspergillus niger HXM2 comprises the following steps:
adding the bacterial liquid containing the fen-flavor Daqu to a starch agar Chaochi culture medium for culturing, and screening to obtain Aspergillus niger HXM2 with the saccharifying enzyme activity of more than 3000 u/g;
the low-temperature Daqu is prepared according to the following method:
making the barley and the peas into curved bricks;
naturally fermenting the bent brick to obtain low-temperature Daqu;
the natural fermentation process comprises the following steps: laying yeast, a mold feeding period, a mold drying period, a moist fire period, a strong fire period, a late fire period and a yeast culturing period, wherein the temperature of the strong fire period is 40-50 ℃;
the yeast is selected from alcoholic yeast HXJ1, and the screening method of the alcoholic yeast HXJ1 comprises the following steps:
inoculating the fen-flavor Daqu in wort of 10-degree Brix, adding lactic acid, mixing, and culturing;
inoculating the culture into 10 ° Brix wort, adding lactic acid, mixing, and culturing;
repeating the steps for 3-4 times to obtain a yeast proliferation solution;
placing the yeast proliferation solution on a wort agar culture medium of 10 DEG Brix for culture, taking a typical colony, inoculating the typical colony on a test tube wort agar slant culture medium of 10 DEG Brix, and then sequentially inoculating the typical colony on the wort agar culture medium of 10 DEG Brix and the test tube wort agar slant culture medium of 10 DEG Brix for culture to obtain an alcohol yeast HXJ1 with the alcohol production performance of more than 4.5% vol;
the ester-producing yeast is selected from ester-producing yeast HXJ3, ester-producing yeast HXJ6 and ester-producing yeast HXJ7, and the screening method of the ester-producing yeast HXJ3, the ester-producing yeast HXJ6 and the ester-producing yeast HXJ7 comprises the following steps:
inoculating the fragrant yeast to wort of 10-degree Brix, adding lactic acid, mixing, and culturing;
inoculating the culture into 10 ° Brix wort, adding lactic acid, mixing, and culturing;
repeating the steps for 3-4 times to obtain a yeast proliferation solution;
placing the yeast proliferation solution on a wort agar culture medium containing streptomycin and having a purity of 10 Brix, culturing, taking a typical bacterial colony, inoculating the typical bacterial colony on a test tube wort agar slant culture medium having a purity of 10 Brix, and then sequentially inoculating the typical bacterial colony on a wort agar culture medium having a purity of 10 Brix and a test tube wort agar slant culture medium having a purity of 10 Brix, and culturing to obtain an ester-producing yeast HXJ3 with an ester-producing property of 4.35g/L of ethyl acetate, an ester-producing yeast HXJ6 with an ester-producing property of 4.20g/L of ethyl acetate, and an ester-producing yeast HXJ7 with an ester-producing property of 3.96g/L of ethyl acetate;
the using amount ratio of the bran koji, the low-temperature Daqu, the yeast and the ester-producing yeast is (3-5): (5-8): (1-3): (1-3);
B) and distilling the fermented grains and storing the distilled liquor to obtain the bran koji fen-flavor liquor.
2. The production method according to claim 1, wherein the ratio of the total inoculation amount of the bran koji, the low-temperature Daqu, the yeast and the ester-producing yeast to the sorghum is (0.10-0.19): 1.
3. the production method according to claim 1, wherein the cellar entry temperature of the sealed fermentation is 14-18 ℃, the cellar entry moisture of the sealed fermentation is 53-60%, the temperature of the sealed fermentation is 20-30 ℃, and the time of the sealed fermentation is 10-15 days.
4. The production method according to claim 1, wherein the distillation is performed as follows:
and (3) uniformly and loosely paving a layer of 30-40 cm fermented grains on the slight strip of the steamer, introducing steam with the pressure of 0.02-0.05MPa, detecting the steam layer by layer, and feeding the steam into the steamer, wherein the liquor flowing speed of each steamer is less than 10 kg per minute, and the liquor flowing temperature is lower than 30 ℃, so that liquor is obtained.
5. The process according to claim 1, wherein the storage time is not less than 120 days.
6. The bran koji fen-flavor liquor obtained by the production method according to any one of claims 1 to 5.
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